Figure S4.

Torin1 induces pexophagy in different cells. (A) Representative fluorescence images of OVCAR8 cells treated with Torin1 (1 μM) or DMSO for 24 h as indicated and immunostained for PMP70. Scale bars: 10 µm. (B) Quantification of peroxisome numbers (green puncta) in cells treated in A. Values are mean ± SD, n = 3 independent experiments, calculated using >90 cells. ***, P < 0.001 by two-tailed Student’s t test. (C) OVCAR8 cells were treated with DMSO or Torin1 for the indicated time. The cell lysates were analyzed by WB and immunoblotted with PMP70 and β-actin antibodies. (D) HeLa cells stably expressing GFP-SKL were treated with DMSO or Torin1 (1 μM) for 24 h. Cells were then fixed and examined by microscopy to detect GFP signal. Scale bars: 5 µm. (E) HeLa cells stably expressing GFP-SKL were treated with or without Torin1 (1 μM for 24 h). Cells were then analyzed by flow cytometry. (F) Quantification of GFP intensity in E. Values are mean ± SD, n = 3 independent experiments. **, P < 0.01; ***, P < 0.001; n.s., not significant by one-way ANOVA and Dunnett’s multiple comparisons test.